mericon Quant RR Soy Kit

For real-time PCR detection and quantification of Roundup Ready Soy DNA in food

Sensitive and specific detection and quantification of Roundup Ready Soy

Harmonized cycling protocol for excellent data comparability

PCR run time of 82 minutes on the QIAGEN Rotor-Gene Q

Part of a complete workflow with dedicated sample preparation kits

Full license for PCR without additional costs

The mericon Quant RR Soy Kit is for the detection and quantification of Roundup Ready soy DNA in food, animal feed, and pharmaceutical products. The assay uses a real-time PCR-based protocol and forms part of the comprehensive QIAGEN food testing portfolio, which also features dedicated sample preparation kits and assays for pathogen detection and ingredient authentication. The mericon Quant RR Soy Kit performs optimally on the Rotor-Gene Q, but has also been validated for block thermal cyclers.

The mericon Quant RR Soy Kit is intended for molecular biology applications in food, animal feed, and pharmaceutical product testing. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Performance

The mericon Quant RR Soy Kit is designed for the specific detection and quantification of Roundup Ready soy DNA in food and animal feed. Both the PCR Assay RR Soy and the PCR Assay Reference System of this kit can detect as few as 10 copies of the target DNA in a reaction.

The PCR Assay RR Soy exhibits high specificity for Roundup Ready soy. In an experiment using 50 ng of tested DNA, no cross-reactivity was observed with other genetic constructs or with plant or animal species not containing this genetic construct (see the table "Results of mericon Quant RR Soy Kit cross-reactivity study").

Results of mericon Quant RR Soy Kit cross-reactivity study

Species, GMO, or genetic construct

Result

Species, GMO, or genetic construct

Result

Roundup Ready soy

+

Bt11 corn

–

Bt176 corn

–

TC1507 corn

–

StarLink corn

–

T25 corn

–

MON810 corn

–

NK603 corn

–

GA21 corn

–

Rice

–

Potato

–

Rye

–

Pig

–

Barley

–

Wheat

–

Sheep

–

The Reference System Assay (soy assay) exhibits high specificity for soy and Roundup ready soy. No cross-reactivity was observed with other plant or animal species or GMOs in an experiment using 50 ng of tested DNA (see the table "Results of Reference System Assay cross-reactivity study").

Results of Reference System Assay cross-reactivity study

Species

Result

Species

Result

Soy

+

Pig

–

Roundup Ready soy

–

Sheep

–

Potato

–

Goat

–

Corn

–

Horse

–

Rape

–

Lupine

–

Rice

–

Hazelnut

–

Barley

–

Pecan nut

–

Wheat

–

Macadamia

–

Rye

–

Almond

–

Cattle

–

Mustard

–

Principle

The mericon Quant Roundup Ready Soy Kit is designed for the quantitative detection of the genetically modified organism (GMO) Roundup Ready soy in food and animal feed samples. The genetic modification in the RR soy line provides resistance to the active compound glyphosate in herbicides, which is detected by the mericon RR Soy Kit. RR soy is the most frequently used genetically modified soy line.

Detecting DNA from a genetically modified organism (GMO) via PCR is based on amplifying a genetic construct used in that GMO. In real-time PCR, the amplified product is detected via target-specific fluorescent probes that bind to the amplified product. Accumulation of PCR product results in an increased fluorescent signal from the bound probes. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating PCR product without having to re-open the reaction tubes afterward.

The probes of mericon PCR Assays are sequence-specific oligonucleotides with a fluorophore at the 5' end of the probe, and a quencher moiety at the 3' end. If the target DNA sequence is present, the probe is cleaved by the 5'→3' exonuclease activity of HotStarTaq Plus DNA Polymerase during the extension phase of PCR. This separates the fluorophore and the quencher moiety, resulting in a detectable fluorescence that is proportional to the amount of accumulated PCR product.

The target GMO is quantified based on the alignment of the PCR results from a GMO-specific PCR system and a Reference PCR system. The GMO assay targets either a genetic construct commonly used in a desired GMO or a DNA sequence that is specific for a particular GMO (e.g., Roundup Ready Soy). The Reference assay targets the plant species of the GMO (e.g., soy). Standard curves for both systems form the basis of the quantification experiment. They are generated with a defined dilution of the Standard DNA, which contains a defined ratio of GMO and Reference DNA.

Each mericon Quant Assay contains an optimized mixture of PCR primer sets for a GMO-specific and a species-specific (Reference System) target sequence, each with an internal control (IC), plus probes labeled with distinct fluorescent dyes (in both systems, the test sample is detected with FAM reporter (495/520 nm), and the internal control is detected with MAX NHS Ester reporter (MAX; 524/557 nm)). In addition, each kit includes a Quantification Control DNA, a Standard DNA, and all of the reagents necessary to perform the analysis.

Procedure

DNA is extracted from a sample of food or animal feed using a dedicated sample preparation kit. QIAGEN's DNeasy mericon Food Kit is suitable for a whole range of food matrices, including those with high inhibitor contents.

The isolated, pure DNA is assayed using a streamlined real-time PCR protocol that returns results in 82 minutes on QIAGEN's Rotor-Gene Q. The protocol has also been validated for use on other thermal block cyclers.